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基质胶:细胞培养最佳生长所需的复杂蛋白质混合物。

Matrigel: a complex protein mixture required for optimal growth of cell culture.

机构信息

Don Rix Protein Identification Facility, Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada.

出版信息

Proteomics. 2010 May;10(9):1886-90. doi: 10.1002/pmic.200900758.

Abstract

Numerous cell types require a surface for attachment to grow and proliferate. Certain cells, particularly primary and stem cells, necessitate the use of specialized growth matrices along with specific culture media conditions to maintain the cells in an undifferentiated state. A gelatinous protein mixture derived from mouse tumor cells and commercialized as Matrigel is commonly used as a basement membrane matrix for stem cells because it retains the stem cells in an undifferentiated state. However, Matrigel is not a well-defined matrix, and therefore can produce a source of variability in experimental results. In this study, we present an in-depth proteomic analysis of Matrigel using a dynamic iterative exclusion method coupled with fractionation protocols that involve ammonium sulfate precipitation, size exclusion chromatography, and one-dimensional SDS-PAGE. The ability to identify the low mass and abundance components of Matrigel illustrates the utility of this method for the analysis of the extracellular matrix, as well as the complexity of the matrix itself.

摘要

许多细胞类型需要附着在表面上才能生长和增殖。某些细胞,特别是原代细胞和干细胞,需要使用专门的生长基质和特定的培养基条件来维持细胞处于未分化状态。一种从老鼠肿瘤细胞中提取的凝胶状蛋白质混合物,商业化后称为 Matrigel,通常被用作干细胞的基底膜基质,因为它能使干细胞保持未分化状态。然而,Matrigel 不是一种明确的基质,因此可能会导致实验结果产生变异性。在这项研究中,我们使用一种动态迭代排除方法结合分馏方案,对 Matrigel 进行了深入的蛋白质组学分析,该方案涉及硫酸铵沉淀、分子筛层析和一维 SDS-PAGE。这种方法能够鉴定 Matrigel 中的低质量和丰度成分,说明了该方法对于分析细胞外基质的有效性,以及基质本身的复杂性。

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